Multiple studies with a single experiment: The Power of ...（二）

A Better Multiplexing Method– Isobaric Mass Tagging

•Less MS1 Complexity

    •Increased Throughput Concurrent MS analysis of multiple samples

    •Less consumed samples and less instrument time

•Fewer Missing Values

    • Identification and quantification achieved in a single run

    •No worries about irreproducibility

•Sample Origin Flexibility

     •Samples can be derived from cells, tissues or biological fluids

•Increased Multiplexing

     •Compare more than 3 conditions

•Multiple Comparisons and Improved Statistics

     •Incorporate replicates with multiple conditions: dose-response, time-course, multiple tissues, subcellular fractions, etc

Thermo Scientific Tandem Mass Tag (TMT) Isobaric Tag Family

*13C and 15N labeled reporter

*Fragments by ETD or HCD

*Isotopes balanced between linker region and reporter region keeping all tags exactly isobaric

*Up to 10 different tags

*Other reactive tags :Iodo TMT and Aminoxy TMT

The Multiplexing Revolution –Not Only Consumables…

High Performance Depends Upon High Resolution Instruments

HIGH RESOLVING POWER IS ESSENTIAL FOR ACCURATE QUANTITATION OF THE TMT10PLEX REAGENTS

Result: Get accurate quantitation using the high resolution of Orbitrap Mass Analyzer

A Real Example

Sample: Mouse mitochondrial extract untreated or treated with phosphatase inhibitor

Orbitrap Elite
 *75 um x 50 cm PepMap C18
 *210 min gradient: 250 min run
 *1 ug of sample on column 

Thermo Poster Note : Liver Mitochondria Proteomics Employing High –Resolution MS Technology; J.Ho. et al